Eu3+-labeled IgG-based time-resolved fluoroimmunoassay for highly sensitive detection of aflatoxin B-1 in feed

文献类型： 外文期刊

第一作者： Zhang, Gaiping

作者： Zhang, Gaiping;Hu, Xiaofei;Yao, Jingjing;Wang, Fangyu;Yin, Mengqi;Sun, Yaning;Hu, Mei;Shi, Qiaoqiao;Zhang, Gaiping

作者机构：

关键词： aflatoxin B-1;monoclonal antibody;europium;time-resolved fluoroimmunoassay

期刊名称：JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE （ 影响因子：3.638； 五年影响因子：3.802 ）

ISSN： 0022-5142

年卷期： 2018 年 98 卷 2 期

页码：

收录情况： SCI

摘要： BACKGROUNDAflatoxin B-1 (AFB(1)) is a kind of toxic and carcinogenic mycotoxin. A time-resolved fluoroimmunoassay (TRFIA) was established for quantitative detection of AFB(1) in feed using Eu3+-labeled IgG as tracer. RESULTSMonoclonal antibody (McAb) against AFB(1) (9B11-D7) was prepared through immunization and cell fusion and was identified as high affinity, specificity and sensibility by enzyme-linked immunosorbent assay (ELISA). The 50% inhibition value (IC50) was 0.81ng mL(-1), the limit of detection (LOD) was 0.10ng mL(-1) and detection range was 0.10-3.94ng mL(-1). Goat anti-mouse immunoglobulin G (IgG) was modified by Eu3+-DATT, generating Eu3+-labeled IgG. Under optimal assay conditions, TRFIA was shown to be highly sensitive and specific in detection of AFB(1). The IC50 and LOD were 94.73pg mL(-1) and 3.55pg mL(-1), respectively, and detection range was 3.55-1.11x10(3)pg mL(-1). Cross-reactivity with AFM(1), AFB(2), AFG(1) and AFG(2) was 31.26%, 37.6%, 127.46% and 35.74%, respectively, but zero with other analogues. In determination of AFB(1) spiked in feed sample, TRFIA showed high accuracy and precision. The average recoveries ranged from 93.71% to 97.80%, and coefficient of variation was 1.25-3.73%. Good correlation between TRFIA and HPLC was demonstrated for determination of AFB(1) in feeds, confirming the reliability of the developed method. CONCLUSIONThe developed TRFIA exhibited good potential for employment in the ultrasensitive detection of AFB(1) in feed and could be used to determine total aflatoxins. (c) 2017 Society of Chemical Industry

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